[Histonet] RE: Antigen retrieval

[C.M. van der Loos]

Kim,How about using a Thermo PTModule for this? You can even program this thing for overnight applications! In the past we used a stove for overnight at 70C. First, a tank with HIER buffer was warmed up in the microwave and then placed in a stove (70C was the maximum setting for it) with a glass plate covering the tank. The effect of overnight antigen retrieval at 70C for the few antigens we tested was a bit different, but mostly comparable with the effect of HIER for 20 min at 98C. Some of the antigens responded very well and showed far more intense staining than after short HIER (20 min, 98C), and others responded very poor. The big advantage of overnight HIER at 70C was that sections from fatty tissue better stayed on the glass slides.Hope this info helps a bit!Cheers, ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands Date: Sat, 12 Apr 2008 17:18:10 +1000
From: Kim O'Sullivan <Kim.Osullivan <@t> med.monash.edu.au>
Subject: [Histonet] Antigen retrieval
To: histonet <@t> lists.utsouthwestern.edu

Hi,

Is anyone familiar with doing antigen retrieval overnight at 80 degrees celsius in 0.1M Tris/HCL pH 9.0? My question is: What is the best way to do this in terms of safety etc. A waterbath? Hot oven? Any idea's appreciated.

Kim O'Sullivan
Monash University 
Melbourne
Australia